# US 12,595,503 B2 · Reference dataset-based, spectrometric characterization of cell substrates using sub-libraries > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 12,595,503 B2 - Document ID: US12595503B2 - Status: Granted - Kind: B2 - Application number: 18/056,107 - Filed: 2022-11-16 - Published: 2023-05-25 - Granted: 2026-04-07 - Earliest claimed: 2021-11-19 - Applicants: Bruker Daltonics GmbH & Co. KG - Inventors: Katrin Sparbier - Web page: https://patentgrep.com/patent/US12595503B2 ## CPC - C12Q 1/10: Enterobacteria - C12Q 1/18: Testing for antimicrobial activity of a material - C12Q 1/04: Determining presence or kind of microorganism; Use of selective media for testing antibiotics or bacteriocides; Compositions containing a chemical indicator therefor {(C12Q1/6897 takes precedence)} - G01N 33/6851: {Methods of protein analysis involving laser desorption ionisation mass spectrometry} - G16B 20/00: ICT specially adapted for functional genomics or proteomics, e.g. genotype-phenotype associations - G16B 40/10: Signal processing, e.g. from mass spectrometry [MS] or from PCR - C12Q: MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS (immunoassay G01N33/53); COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES - G01N: INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES (measuring or testing apparatus or processes other than immunoassay, involving enzymes or microorganisms C12M, C12Q) - G16B: BIOINFORMATICS, i.e. INFORMATION AND COMMUNICATION TECHNOLOGY [ICT] SPECIALLY ADAPTED FOR GENETIC OR PROTEIN-RELATED DATA PROCESSING IN COMPUTATIONAL MOLECULAR BIOLOGY ## Abstract The invention relates to methods for spectrometric characterization of a test cell substrate. The characterization comprises taxonomic classification and determination of a property of interest of the test cell substrate. The characterization may be based on mass-spectrometric measurement data. The property of interest may be a resistance or susceptibility to a growth-influencing factor. After comparing first spectrometric measurement data of the test cell substrate with a provided reference library, a sub-library is created comprising those reference datasets from the reference library that are classified as allowing a taxonomic classification of the test cell substrate. Second spectrometric measurement data after a second preparation of the test cell substrate under conditions that serve to determine a property of interest of the test cell substrate is compared with the sub-library and allow a reliable determination of the property of interest. ## Claims ### Claim 1 A method for spectrometrically characterizing a test cell substrate, comprising: a) providing a library comprising a multitude of reference data sets, with each reference data set containing data that allows a taxonomic classification of a cell substrate; b) obtaining first spectrometric measurement data from the test cell substrate after a first preparation; c) comparing the first spectrometric measurement data or data derived therefrom with the library to determine a first match result, wherein a match result contains a list of reference data sets and their degree of matching with spectrometric measurement data of a cell substrate or data derived therefrom; d) creating a sub-library comprising reference data sets from the library for which the first match result is evaluated as allowing a taxonomic classification of the test cell substrate; e) obtaining second spectrometric measurement data from the test cell substrate after at least a second preparation under conditions that are not identical to the conditions of the first preparation; f) comparing the second spectrometric measurement data or data derived therefrom with the sub-library to determine a second match result; and g) determining a property of the test cell substrate using the second match result. ### Claim 2 (depends on 1) The method according to claim 1, wherein the taxonomic classification comprises assignment to the test cell substrate of a taxon selected from the group: genus; species; subspecies; and variety or serotype. ### Claim 3 (depends on 1) The method according claim 1, wherein the test cell substrate comprises bacteria of the family Enterobacteriaceae. ### Claim 4 (depends on 1) The method according to claim 1, wherein each of the first preparation and second preparation include a multiplication step of the test cell substrate. ### Claim 5 (depends on 1) The method according to claim 1, wherein the second preparation is carried out in the presence of a growth-influencing factor. ### Claim 6 (depends on 1) The method according to claim 1, wherein the first preparation is carried out as a growth control without using a growth-influencing factor. ### Claim 7 (depends on 1) The method according to claim 1, wherein a reference data set comprises a spectrum or a data n-tuple derived from a spectrum. ### Claim 8 (depends on 1) The method according to claim 1, wherein the property to be determined comprises a susceptibility and/or resistance of the test cell substrate to a growth-influencing factor. ### Claim 9 (depends on 8) The method according to claim 8, wherein the property to be determined comprises a minimum inhibitory concentration (MIC) of the growth-influencing factor for the test cell substrate. ### Claim 10 (depends on 1) The method according to claim 1, wherein reference data sets for which the first match result is evaluated as not allowing a taxonomic classification of the test cell substrate are excluded from the creation of the sub-library. ### Claim 11 (depends on 1) The method according to claim 1, wherein the sub-library is a first sub-library, and wherein a second sub-library is created that comprises reference data sets from the library for which the first match result is evaluated as not allowing a taxonomic classification of the test cell substrate, and wherein the second spectrometric measurement data or data derived therefrom is compared with the second sub-library in order to determine a third match result, and a property of the test cell substrate is determined using the third match result. ### Claim 12 (depends on 1) The method according to claim 1, wherein the obtaining of the first and/or the second spectrometric measurement data from the test cell substrate comprises mass spectrometric analysis. ### Claim 13 (depends on 1) The method according to claim 1, further comprising further preparations of the test cell substrate under conditions that are not identical to the conditions of the first preparation or second preparation or with each other. ### Claim 14 (depends on 1) The method according to claim 1, wherein the first preparation and/or the second preparation is carried out directly on a sample support which serves as a substrate for obtaining spectrometric measurement data. ### Claim 15 (depends on 1) The method according to claim 1, wherein: a) the test cell substrate comprises bacteria of the family Enterobacteriaceae; b) the first preparation and second preparation each comprise a multiplication step, c) the second preparation is carried out in the presence of a growth-influencing factor; d) the first preparation is carried out as a growth control without the use of a growth-influencing factor; e) the taxonomic classification comprises assignment to the test cell substrate of a taxon selected from the group consisting of: genus; species; subspecies; and variety or serotype; f) at least one of the reference data sets comprises a spectrum or a data n-tuple derived from a spectrum; g) the growth-influencing factor is an antimicrobial agent, and the property of the test cell substrate is a susceptibility or resistance of the test cell substrate to the antimicrobial agent; h) reference data sets for which the first match result is evaluated as not allowing a taxonomic classification of the test cell substrate are excluded in the provision or creation of the sub-library; i) the sub-library is a first sub-library, and wherein a second sub-library is created that comprises the reference data sets from the library for which the first match result is evaluated as not allowing a taxonomic classification of the test cell substrate, and wherein the second spectrometric measurement data or data derived therefrom is compared with the second sub-library to determine a third match result; j) the third match result is evaluated as not allowing taxonomic classification of the test cell substrate; and k) the obtaining of first and/or second spectrometric measurement data from the test cell substrate comprises a mass spectrometric analysis.